Effect of aging and oral tolerance on dendritic cell function

Oral tolerance can be induced in some mouse strains by gavage or spontaneous ingestion of dietary antigens. In the present study, we determined the influence of aging and oral tolerance on the secretion of co-stimulatory molecules by dendritic cells (DC), and on the ability of DC to induce proliferation and cytokine secretion by naive T cells from BALB/c and OVA transgenic (DO11.10) mice. We observed that oral tolerance could be induced in BALB/c mice (N = 5 in each group) of all ages (8, 20, 40, 60, and 80 weeks old), although a decline in specific antibody levels was observed in the sera of both tolerized and immunized mice with advancing age (40 to 80 weeks old). DC obtained from young, adult and middle-aged (8, 20, and 40 weeks old) tolerized mice were less efficient (65, 17 and 20%, respectively) than DC from immunized mice (P < 0.05) in inducing antigen-specific proliferation of naive T cells from both BALB/c and DO11.10 young mice, or in stimulating IFN-g, IL-4 and IL-10 production. However, TGF-&#946; levels were significantly elevated in co-cultures carried out with DC from tolerant mice (P < 0.05). DC from both immunized and tolerized old and very old (60 and 80 weeks old) mice were equally ineffective in inducing T cell proliferation and cytokine production (P < 0.05). A marked reduction in CD86+ marker expression was observed in DC isolated from both old and tolerized mice (75 and 50%, respectively). The results indicate that the aging process does not interfere with the establishment of oral tolerance in BALB/c mice, but reduces DC functions, probably due to the decline of the expression of the CD86 surface marker.687

Aging alters the production of iNOS, arginase and cytokines in murine macrophages

The limited amount of information on the primary age-related deficiencies in the innate immune system led us to study the production of inducible nitric oxide synthase (iNOS), arginase, and cytokines in macrophages of young (8 weeks old) and old (72 weeks old) female BALB/c mice. We first evaluated iNOS and arginase inducers on peritoneal (PM&#934;) and bone marrow-derived (BMM&#934;) macrophages of young BALB/c and C57BL/6 mice, and then investigated their effects on macrophages of old mice. Upon stimulation with lipopolysaccharide (LPS), resident and thioglycolate-elicited PM&#934; from young mice presented higher iNOS activity than those from old mice (54.4%). However, LPS-stimulated BMM&#934; from old mice showed the highest NO levels (50.1%). Identical NO levels were produced by PM&#934; and BMM&#934; of both young and old mice stimulated with interferon-&#947;. Arginase activity was higher in resident and elicited PM&#934; of young mice stimulated with LPS (48.8 and 32.7%, respectively) and in resident PM&#934; stimulated with interleukin (IL)-4 (64%). BMM&#934; of old mice, however, showed higher arginase activity after treatment with IL-4 (46.5%). In response to LPS, PM&#934; from old mice showed the highest levels of IL-1&#945; (772.3 ± 51.9 pg/mL), whereas, those from young mice produced the highest amounts of tumor necrosis factor (TNF)-&#945; (937.2 ± 132.1 pg/mL). Only TNF-&#945; was expressed in LPS-treated BMM&#934;, and cells from old mice showed the highest levels of this cytokine (994.1 ± 49.42 pg/mL). Overall, these results suggest that macrophages from young and old mice respond differently to inflammatory stimuli, depending on the source and maturity of the cell donors.67168

Aging reduces the primary humoral response and the in vitro cytokine production in mice

Aging is accompanied by a decrease in several physiological functions that make older individuals less responsive to environmental challenges. In the present study, we analyzed the immune response of female BALB/c mice (N = 6) of different ages (from 2 to 96 weeks) and identified significant age-related alterations. Immunization with hapten-protein (trinitrophenyl-bovine serum albumin) conjugates resulted in lower antibody levels in the primary and secondary responses of old mice (72 weeks old). Moreover, young mice (2, 16, and 32 weeks old) maintained specific antibodies in their sera for longer periods after primary immunization than did old mice. However, a secondary challenge efficiently induced memory in old mice, as shown by the increased antibody levels in their sera. The number of CD4+ and CD8+ T cells in the spleen increased until 8 weeks of age but there was no change in the CD4+/CD8+ ratio with aging. Splenic T cells from old mice that had or had not been immunized were less responsive to concanavalin-A and showed reduced cytokine production compared to young mice (IL-2: 57-127 vs 367-1104 pg/mL, IFN-g: 2344-12,836 vs 752-23,106 pg/mL and IL-10: 393-2172 vs 105-2869 pg/mL in old and young mice, respectively). These data suggest that there are significant changes in the organization of the immune system throughout life. However, the relevance of these alterations for the functioning of the immune system is unknown.1111112

Aging reduces the primary humoral response and the in vitro cytokine production in mice

Aging is accompanied by a decrease in several physiological functions that make older individuals less responsive to environmental challenges. In the present study, we analyzed the immune response of female BALB/c mice (N = 6) of different ages (from 2 to 96 weeks) and identified significant age-related alterations. Immunization with hapten-protein (trinitrophenyl-bovine serum albumin) conjugates resulted in lower antibody levels in the primary and secondary responses of old mice (72 weeks old). Moreover, young mice (2, 16, and 32 weeks old) maintained specific antibodies in their sera for longer periods after primary immunization than did old mice. However, a secondary challenge efficiently induced memory in old mice, as shown by the increased antibody levels in their sera. The number of CD4+ and CD8+ T cells in the spleen increased until 8 weeks of age but there was no change in the CD4+/CD8+ ratio with aging. Splenic T cells from old mice that had or had not been immunized were less responsive to concanavalin-A and showed reduced cytokine production compared to young mice (IL-2: 57-127 vs 367-1104 pg/mL, IFN-g: 2344-12,836 vs 752-23,106 pg/mL and IL-10: 393-2172 vs 105-2869 pg/mL in old and young mice, respectively). These data suggest that there are significant changes in the organization of the immune system throughout life. However, the relevance of these alterations for the functioning of the immune system is unknown

Effect of aging and oral tolerance on dendritic cell function

Oral tolerance can be induced in some mouse strains by gavage or spontaneous ingestion of dietary antigens. In the present study, we determined the influence of aging and oral tolerance on the secretion of co-stimulatory molecules by dendritic cells (DC), and on the ability of DC to induce proliferation and cytokine secretion by naive T cells from BALB/c and OVA transgenic (DO11.10) mice. We observed that oral tolerance could be induced in BALB/c mice (N = 5 in each group) of all ages (8, 20, 40, 60, and 80 weeks old), although a decline in specific antibody levels was observed in the sera of both tolerized and immunized mice with advancing age (40 to 80 weeks old). DC obtained from young, adult and middle-aged (8, 20, and 40 weeks old) tolerized mice were less efficient (65, 17 and 20%, respectively) than DC from immunized mice (P < 0.05) in inducing antigen-specific proliferation of naive T cells from both BALB/c and DO11.10 young mice, or in stimulating IFN-g, IL-4 and IL-10 production. However, TGF-&#946; levels were significantly elevated in co-cultures carried out with DC from tolerant mice (P < 0.05). DC from both immunized and tolerized old and very old (60 and 80 weeks old) mice were equally ineffective in inducing T cell proliferation and cytokine production (P < 0.05). A marked reduction in CD86+ marker expression was observed in DC isolated from both old and tolerized mice (75 and 50%, respectively). The results indicate that the aging process does not interfere with the establishment of oral tolerance in BALB/c mice, but reduces DC functions, probably due to the decline of the expression of the CD86 surface marker

Parallels Between Plants And Animals In The Production And Molecular Targets Of Nitric Oxide

Considerable evidence that nitric oxide (NO) and its derivatives play major roles in mammals has led to an interest in the actions of these molecules in plant metabolism. The ubiquitous distribution of nitric oxide synthases (NOS) in mammalian cells has stimulated the search for an equivalent enzyme in plants. NOS-like activity has been found in many plants and NO has been shown to influence various developmental processes and to have a role in plant defense responses against pathogens. Several of the major NO targets characterized in animals also have found similar actions in plants. 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Aging alters the production of iNOS, arginase and cytokines in murine macrophages

The limited amount of information on the primary age-related deficiencies in the innate immune system led us to study the production of inducible nitric oxide synthase (iNOS), arginase, and cytokines in macrophages of young (8 weeks old) and old (72 weeks old) female BALB/c mice. We first evaluated iNOS and arginase inducers on peritoneal (PMΦ) and bone marrow-derived (BMMΦ) macrophages of young BALB/c and C57BL/6 mice, and then investigated their effects on macrophages of old mice. Upon stimulation with lipopolysaccharide (LPS), resident and thioglycolate-elicited PMΦ from young mice presented higher iNOS activity than those from old mice (54.4%). However, LPS-stimulated BMMΦ from old mice showed the highest NO levels (50.1%). Identical NO levels were produced by PMΦ and BMMΦ of both young and old mice stimulated with interferon-γ. Arginase activity was higher in resident and elicited PMΦ of young mice stimulated with LPS (48.8 and 32.7%, respectively) and in resident PMΦ stimulated with interleukin (IL)-4 (64%). BMMΦ of old mice, however, showed higher arginase activity after treatment with IL-4 (46.5%). In response to LPS, PMΦ from old mice showed the highest levels of IL-1α (772.3 ± 51.9 pg/mL), whereas, those from young mice produced the highest amounts of tumor necrosis factor (TNF)-α (937.2 ± 132.1 pg/mL). Only TNF-α was expressed in LPS-treated BMMΦ, and cells from old mice showed the highest levels of this cytokine (994.1 ± 49.42 pg/mL). Overall, these results suggest that macrophages from young and old mice respond differently to inflammatory stimuli, depending on the source and maturity of the cell donors

Kinetic Modeling Of Cell Death By Shear Stress Of A Hybridoma Culture In A Sparged And Stirred Bioreactor

In animal cell systems, cell death by shear stress is an important factor to be considered when defining the required sparging and agitation to maintain the dissolved oxygen concentration and the homogeneity of the bioreactor. This paper presents the effects of air sparging and agitation in an ideal bioreactor configuration for animal cell culture. In these conditions, the effect of agitation on the cell death rate is negligible and this parameter can be considered directly proportional to the superficial gas rate.49SPEC. ISS.107114Abe, K., Matsuki, N., Measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- tetrazolium bromide (MTT) reduction activity and lactate dehydrogenase release using MTT (2000) Neuroscience Research, 38, pp. 325-329Aloi, L.E., Cherry, R.S., Cellular response to agitation characterized by energy Dissipation at the impeller tip (1996) Chem. Engng Sci., 51 (9), pp. 1523-1529Apenberg, S., Freyberg, M.A., Friedl, P., Shear stress induces apoptosis in vascular smooth muscle cells via an autocrine Fas/FasL pathway (2003) Biochem. Biophys. Res. Commun., 301, pp. 355-359Aunins, J.G., Henzler, H.J., Aeration in cell culture bioreactors (1993) BiotechnologyA Multivolume Treatise, 2nd Edition, pp. 219-281. , eds. Rehm, H. J.Reed, G., Federal Republic of Germany, VCHBliem, R., Katinger, H., Scale-up engineering in animal cell technology: Part II (1988) Tibtech, 6, pp. 224-230Butler, M., The characteristics and growth of cultured cells (1991) Mammalian Cell Biotechnology: A Practical Approach, pp. 1-25. , ed. Butler, M., Great Britain, IRL PressChisti, Y., Animal cell damage in sparged bioreactors (2000) Tibtech, 18, pp. 420-432Chist, Y., Hydrodynamic damage to animal cells (2001) Critical Reviews in Biotechnology, 21 (2), pp. 67-110Feuser, J., Halfar, M., Lütkemeyer, Ameskamp, N., Kula, M.-R., Thömmes, J., Interaction of mammalian cell culture broth with adsorbents in expanded bed adsorption of monoclonal antibodies (1999) Process Biochemistry, 34, pp. 159-165Gardner, A.R., Gainer, J.L., Kirwan, D.J., Effects of stirring and sparging on cultured hybridoma cells (1990) Biotechnol Bioeng., 35, pp. 940-947Handa-Corrigan, A., Emery, A.N., Spier, R.E., Effect of gas-liquid interfaces on the growth of suspended mammalian cells: Mechanisms of cell damage by bubbles (1989) Enzyme Microb. Technol., 11, pp. 230-235Kunas, T.K., Papoutsakis, E.T., Damage mechanisms of suspended animal cells in agitated bioreactors with and without bubble entrainment (1990) Biotechnol. Bioeng., 36, pp. 476-483Lash, L.H., Tokarz, J.J., Pegouske, D.M., Susceptibility of primary cultures of proximal tubular and distal tubular cells from rat kidney to chemical induced toxicity (1995) Toxicology, 103, pp. 85-103Leist, C.H., Meyer, H.P., Fiechter, A., A potential and problems of animal cells in suspension (1990) J. Biotechnol., 15, pp. 1-46Léo, P., Ucelli, P., Augusto, E.F.P., Oliveira, M.S., Tamashiro, W.M.S.C., Anti-TNP monoclonal antibodies as reagents for enzyme immunoassay (ELISA) (2000) Hybridoma, 19 (6), pp. 473-479Li, C., Xu, Q., Mechanical stress-initiated signal transductions in vascular smooth muscle cells (2000) Cellular Signalling, 12, pp. 435-445Mardikar, S.H., Niranjan, K., Observations on shear damage to different animal cells in a concentric cylinder viscometer (2000) Biotechnol. Bioeng., 68 (6), pp. 697-704Neter, J., Kutner, M.H., Nachtsheim, C.J., Wasserman, W., (1996) Applied Linear Statistical Models, 4th Edition, pp. 217-241. , WCB/MacGraw-HillPapoutsakis, E.T., Fluid-mechanical damage of animal cells in bioreactors (1991) Tibtech, 9, pp. 427-437Tramper, J., Vlak, J.M., Gooijer, C.D., Scale-up aspects of sparged insect cells bioreactors (1996) Cytotechnology, 20, pp. 221-229Varley, J., Birch, J., Reactor design for large scale suspension animal cell culture (1999) Cytotechnology, 29, pp. 177-205Wagner, A., Marc, A., Engasser, J.M., Einsele, A., The use of Lactate Dehydrogenase (LDH) release kinetics for the evaluation of death and growth of mammalian cells in perfusion reactors (1992) Biotechnol. Bioeng., 39, pp. 320-326Wu, J., Mechanisms of animal cell damage associated with gas bubbles and cell protection by medium additives (1995) J. Biotechnol., 43, pp. 81-9